Rapid screening of pKa values of pharmaceuticals by pressure-assisted capillary electrophoresis combined with short-end injection

A method applying pressure-assisted capillary electrophoresis combined with short-end injection has been developed for the rapid screening of the pk(a) values of pharmaceuticals. The electrophoretic separation is performed on a short capillary length with short-end injection under an applied pressure, and the effective mobility is measured in a series of 10 different buffers with constant ionic strength (I = 0.05). The application of pressure not only reduces migration times, particularly in lower pH buffers, but also improves the repeatability of effective mobility measurements. The influence of pressure on the effective mobility was investigated at various pH values. It was observed for the first time that an increase in pressure resulted in a slight decrease in the effective mobility when the pH was above the pK(a) for acidic analytes, whereas an increased effective mobility with increasing pressures was observed when the pH was below the pK(a). However, the observed effective mobility shift by the applied pressure did not significantly affect the determined pK(a) values. The determined pK(a) values were in good agreement with published data. Furthermore, a stacking condition was applied to increase the sensitivity, and a concentration down to 2 muM could readily be detected with UV detection using a 50 mum I.D. capillary. This technique is particularly suitable for measurement of pK(a) values for compounds with poor aqueous solubility. The method also omits the commonly used preconditioning steps with sodium hydroxide and water. The exclusion of excessive preconditioning steps and the use of pressure reduces the total cycling analysis time, and makes it possible to determine the pK(a) in less than 40 min per compound without loss of accuracy. (C) 2002 Elsevier Science B.V. All rights reserved.